Development of a neutralizing mouse-pig chimeric antibody with therapeutic potential against Haemophilus parasuis in Pichia pastoris

FEMS Microbiol Lett. 2014 May;354(2):85-91. doi: 10.1111/1574-6968.12437. Epub 2014 Apr 16.

Abstract

Haemophilus parasuis is one of the most important bacterial diseases of pigs worldwide. The lack of a vaccine against a broad spectrum of strains and the limitation of antimicrobial susceptibility hamper the control of disease. In this study, we cloned the constant regions of gamma heavy chains and kappa light chain of pig lymphocytes in frame with the variable regions of heavy and light chains of mouse monoclonal antibody 1D8, which reacts with all 15 serotypes of H. parasuis and has neutralizing activity. The constructed mouse-pig chimeric antibody was expressed in Pichia pastoris. Results demonstrated that the expressed chimeric antibody inhibited the growth of H. parasuis in vitro. Furthermore, the experiments in mice showed that chimeric antibody increased survival rate of the mice compared with that of the control group (P < 0.05). Importantly, the chimeric antibody partially protected piglets against H. parasuis infection according to the clinical lesion scores and PCR results of H. parasuis in the tissues from piglets of the chimeric antibody-inoculated group and the PBS group. In summary, our results demonstrated that the mouse-pig chimeric antibody could be a therapeutic candidate to prevent the H. parasuis infection and control the prevalence of disease.

Keywords: Haemophilus parasuis; Pichia pastoris; chimeric antibody; immunotherapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bacterial / genetics*
  • Antibodies, Bacterial / immunology
  • Antibodies, Bacterial / therapeutic use
  • Antibodies, Monoclonal / genetics*
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / therapeutic use
  • Antibodies, Neutralizing / genetics*
  • Antibodies, Neutralizing / immunology
  • Antibodies, Neutralizing / therapeutic use
  • Gene Expression
  • Haemophilus Infections / drug therapy
  • Haemophilus Infections / microbiology
  • Haemophilus Infections / veterinary*
  • Haemophilus parasuis / drug effects
  • Haemophilus parasuis / genetics
  • Haemophilus parasuis / growth & development
  • Haemophilus parasuis / immunology*
  • Mice
  • Mice, Inbred BALB C
  • Pichia / genetics*
  • Pichia / metabolism
  • Protein Engineering
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / therapeutic use
  • Swine
  • Swine Diseases / drug therapy*
  • Swine Diseases / microbiology
  • Swine Diseases / prevention & control

Substances

  • Antibodies, Bacterial
  • Antibodies, Monoclonal
  • Antibodies, Neutralizing
  • Recombinant Fusion Proteins